Figure 3: Efficiency and fidelity of OEPR cloning for different sizes of homology arms.
From: OEPR Cloning: an Efficient and Seamless Cloning Strategy for Large- and Multi-Fragments
(A) Different homology arms (5, 10, 15, 20, 25, 30 and 35âbp (lanes 1â7, respectively)) were added to the 1âkb fragment from hNaV1.5. PCR product were detected by agarose gel electrophoresis. The arrow indicates the target band of the first PCR. (B) The purified fragments (containing 5, 10, 15, 20, 25, 30 and 35âbp homology arms (lanes 8â14, respectively)) from the 1st PCR were inserted into pGADT7 (7988âbp). PCR products were detected by agarose gel electrophoresis. The arrow indicates the target band of the second PCR. (C) Colony numbers per plate were counted to estimate cloning efficiencies. (D) The percentage of positive clones, which was estimated using colony PCR, was used to estimate cloning fidelities. Reported results are the meanâÂąâSEM of three independent experiments. M: DNA molecular weight marker.
